{
"METABOLOMICS WORKBENCH":{"STUDY_ID":"ST002490","ANALYSIS_ID":"AN004064","VERSION":"1","CREATED_ON":"February 21, 2023, 12:40 pm"},

"PROJECT":{"PROJECT_TITLE":"Simultaneous targeting of PD-1 and IL2Rβγ with radiation therapy to inhibit pancreatic cancer growth and metastasis","PROJECT_SUMMARY":"In pancreatic ductal adenocarcinoma (PDAC) patients, we show that response to radiation therapy (RT) is characterized by increased IL2Rβ and IL2Rγ and decreased ILR2α expression. The bispecific aPD1-IL2v is a PD-1-targeted IL-2 variant (IL2v) immunocytokine with engineered IL-2 cis-targeted to PD-1 and abolished IL2Rα binding, which enhances tumor-antigen specific T cell activation while reducing regulatory T cell (Treg) suppression. Using aPD1-IL2v in orthotopic PDAC KPC-driven tumor models, we show marked improvement in local and metastatic survival along with profound increase in tumor-infiltrating polyfunctional CD8+ T cell subsets with a transcriptionally and metabolically active phenotype, and preferential activation of antigen-specific CD8+ T cells. In combination with single dose RT, aPD1-IL2v treatment results in a robust, durable expansion of polyfunctional CD8+ T cells, T cell stemness, tumor-specific memory immune response, natural killer (NK) cell activation, and decreased Tregs. These data show that aPD1-IL2v leads to profound local and distant response in PDAC.","INSTITUTE":"University of Colorado Denver","LABORATORY":"Lab of Angelo D'Alessandro in collaboration with lab of Sana Karam","LAST_NAME":"Haines","FIRST_NAME":"Julie","ADDRESS":"12801 E 17th Ave, Room 1303, Aurora, Colorado, 80045, USA","EMAIL":"julie.haines@cuanschutz.edu","PHONE":"3037243339"},

"STUDY":{"STUDY_TITLE":"Simultaneous targeting of PD-1 and IL2Rβγ with radiation therapy to inhibit pancreatic cancer growth and metastasis - human plasma metabolomics","STUDY_SUMMARY":"Patient plasma samples were collected as part of a Phase I radiation dose-escalation clinical trial (NCT02873598) before, during (6 hours post), and post (6 weeks) SBRT. Samples were collected and analyzed per COMIRB19–0328 for branched chain amino acids and kynurenine.","INSTITUTE":"University of Colorado Denver","LAST_NAME":"Haines","FIRST_NAME":"Julie","ADDRESS":"12801 E 17th Ave, Room 1303, Aurora, Colorado, 80045, USA","EMAIL":"julie.haines@cuanschutz.edu","PHONE":"3037243339"},

"SUBJECT":{"SUBJECT_TYPE":"Human","SUBJECT_SPECIES":"Homo sapiens","TAXONOMY_ID":"9606"},
"SUBJECT_SAMPLE_FACTORS":[
{
"Subject ID":"-",
"Sample ID":"1",
"Factors":{"Patient ID":"C04","Timepoint":"Baseline"},
"Additional sample data":{"RAW_FILE_NAME":"BW49-01_r18"}
},
{
"Subject ID":"-",
"Sample ID":"4",
"Factors":{"Patient ID":"C05","Timepoint":"Baseline"},
"Additional sample data":{"RAW_FILE_NAME":"BW49-02_r11"}
},
{
"Subject ID":"-",
"Sample ID":"7",
"Factors":{"Patient ID":"C06","Timepoint":"Baseline"},
"Additional sample data":{"RAW_FILE_NAME":"BW49-03_r5"}
},
{
"Subject ID":"-",
"Sample ID":"10",
"Factors":{"Patient ID":"C08","Timepoint":"Baseline"},
"Additional sample data":{"RAW_FILE_NAME":"BW49-04_r33"}
},
{
"Subject ID":"-",
"Sample ID":"13",
"Factors":{"Patient ID":"C09","Timepoint":"Baseline"},
"Additional sample data":{"RAW_FILE_NAME":"BW49-05_r9"}
},
{
"Subject ID":"-",
"Sample ID":"16",
"Factors":{"Patient ID":"C10","Timepoint":"Baseline"},
"Additional sample data":{"RAW_FILE_NAME":"BW49-06_r22"}
},
{
"Subject ID":"-",
"Sample ID":"19",
"Factors":{"Patient ID":"C11","Timepoint":"Baseline"},
"Additional sample data":{"RAW_FILE_NAME":"BW49-07_r31"}
},
{
"Subject ID":"-",
"Sample ID":"22",
"Factors":{"Patient ID":"C13","Timepoint":"Baseline"},
"Additional sample data":{"RAW_FILE_NAME":"BW49-08_r13"}
},
{
"Subject ID":"-",
"Sample ID":"25",
"Factors":{"Patient ID":"C14","Timepoint":"Baseline"},
"Additional sample data":{"RAW_FILE_NAME":"BW49-09_r35"}
},
{
"Subject ID":"-",
"Sample ID":"28",
"Factors":{"Patient ID":"C15","Timepoint":"Baseline"},
"Additional sample data":{"RAW_FILE_NAME":"BW49-10_r3"}
},
{
"Subject ID":"-",
"Sample ID":"31",
"Factors":{"Patient ID":"C16","Timepoint":"Baseline"},
"Additional sample data":{"RAW_FILE_NAME":"BW49-11_r1"}
},
{
"Subject ID":"-",
"Sample ID":"34",
"Factors":{"Patient ID":"C17","Timepoint":"Baseline"},
"Additional sample data":{"RAW_FILE_NAME":"BW49-12_r23"}
},
{
"Subject ID":"-",
"Sample ID":"2",
"Factors":{"Patient ID":"C04","Timepoint":"During"},
"Additional sample data":{"RAW_FILE_NAME":"BW49-13_r14"}
},
{
"Subject ID":"-",
"Sample ID":"5",
"Factors":{"Patient ID":"C05","Timepoint":"During"},
"Additional sample data":{"RAW_FILE_NAME":"BW49-14_r28"}
},
{
"Subject ID":"-",
"Sample ID":"8",
"Factors":{"Patient ID":"C06","Timepoint":"During"},
"Additional sample data":{"RAW_FILE_NAME":"BW49-15_r21"}
},
{
"Subject ID":"-",
"Sample ID":"11",
"Factors":{"Patient ID":"C08","Timepoint":"During"},
"Additional sample data":{"RAW_FILE_NAME":"BW49-16_r7"}
},
{
"Subject ID":"-",
"Sample ID":"14",
"Factors":{"Patient ID":"C09","Timepoint":"During"},
"Additional sample data":{"RAW_FILE_NAME":"BW49-17_r12"}
},
{
"Subject ID":"-",
"Sample ID":"17",
"Factors":{"Patient ID":"C10","Timepoint":"During"},
"Additional sample data":{"RAW_FILE_NAME":"BW49-18_r30"}
},
{
"Subject ID":"-",
"Sample ID":"20",
"Factors":{"Patient ID":"C11","Timepoint":"During"},
"Additional sample data":{"RAW_FILE_NAME":"BW49-19_r10"}
},
{
"Subject ID":"-",
"Sample ID":"23",
"Factors":{"Patient ID":"C13","Timepoint":"During"},
"Additional sample data":{"RAW_FILE_NAME":"BW49-20_r4"}
},
{
"Subject ID":"-",
"Sample ID":"26",
"Factors":{"Patient ID":"C14","Timepoint":"During"},
"Additional sample data":{"RAW_FILE_NAME":"BW49-21_r32"}
},
{
"Subject ID":"-",
"Sample ID":"29",
"Factors":{"Patient ID":"C15","Timepoint":"During"},
"Additional sample data":{"RAW_FILE_NAME":"BW49-22_r26"}
},
{
"Subject ID":"-",
"Sample ID":"32",
"Factors":{"Patient ID":"C16","Timepoint":"During"},
"Additional sample data":{"RAW_FILE_NAME":"BW49-23_r29"}
},
{
"Subject ID":"-",
"Sample ID":"35",
"Factors":{"Patient ID":"C17","Timepoint":"During"},
"Additional sample data":{"RAW_FILE_NAME":"BW49-24_r15"}
},
{
"Subject ID":"-",
"Sample ID":"3",
"Factors":{"Patient ID":"C04","Timepoint":"Post"},
"Additional sample data":{"RAW_FILE_NAME":"BW49-25_r6"}
},
{
"Subject ID":"-",
"Sample ID":"6",
"Factors":{"Patient ID":"C05","Timepoint":"Post"},
"Additional sample data":{"RAW_FILE_NAME":"BW49-26_r36"}
},
{
"Subject ID":"-",
"Sample ID":"9",
"Factors":{"Patient ID":"C06","Timepoint":"Post"},
"Additional sample data":{"RAW_FILE_NAME":"BW49-27_r20"}
},
{
"Subject ID":"-",
"Sample ID":"12",
"Factors":{"Patient ID":"C08","Timepoint":"Post"},
"Additional sample data":{"RAW_FILE_NAME":"BW49-28_r2"}
},
{
"Subject ID":"-",
"Sample ID":"15",
"Factors":{"Patient ID":"C09","Timepoint":"Post"},
"Additional sample data":{"RAW_FILE_NAME":"BW49-29_r19"}
},
{
"Subject ID":"-",
"Sample ID":"18",
"Factors":{"Patient ID":"C10","Timepoint":"Post"},
"Additional sample data":{"RAW_FILE_NAME":"BW49-30_r8"}
},
{
"Subject ID":"-",
"Sample ID":"21",
"Factors":{"Patient ID":"C11","Timepoint":"Post"},
"Additional sample data":{"RAW_FILE_NAME":"BW49-31_r24"}
},
{
"Subject ID":"-",
"Sample ID":"24",
"Factors":{"Patient ID":"C13","Timepoint":"Post"},
"Additional sample data":{"RAW_FILE_NAME":"BW49-32_r27"}
},
{
"Subject ID":"-",
"Sample ID":"27",
"Factors":{"Patient ID":"C14","Timepoint":"Post"},
"Additional sample data":{"RAW_FILE_NAME":"BW49-33_r25"}
},
{
"Subject ID":"-",
"Sample ID":"30",
"Factors":{"Patient ID":"C15","Timepoint":"Post"},
"Additional sample data":{"RAW_FILE_NAME":"BW49-34_r24"}
},
{
"Subject ID":"-",
"Sample ID":"33",
"Factors":{"Patient ID":"C16","Timepoint":"Post"},
"Additional sample data":{"RAW_FILE_NAME":"BW49-35_r17"}
}
],
"COLLECTION":{"COLLECTION_SUMMARY":"Patient plasma samples were collected as part of a Phase I radiation dose-escalation clinical trial (NCT02873598) before, during (6 hours post), and post (6 weeks) SBRT. Samples were collected and analyzed per COMIRB19–0328.","SAMPLE_TYPE":"Blood (plasma)"},

"TREATMENT":{"TREATMENT_SUMMARY":"All patients received neoadjuvant chemotherapy prior to SBRT. SBRT doses ranged from 9Gy × 3 fractions to 11Gy ×3 fractions."},

"SAMPLEPREP":{"SAMPLEPREP_SUMMARY":"Plasma aliquots were thawed on ice then a 20 uL aliquot treated with 480 uL of cold 5:3:2 MeOH:acetonitrile:water. Samples were vortexed 30 min at 4 degrees C then supernatants clarified by centrifugation (10 min, 10,000 g, 4 degrees C) and transferred to autosampler vials.","PROCESSING_STORAGE_CONDITIONS":"4℃","EXTRACT_STORAGE":"-80℃"},

"CHROMATOGRAPHY":{"CHROMATOGRAPHY_SUMMARY":"Positive C18","CHROMATOGRAPHY_TYPE":"Reversed phase","INSTRUMENT_NAME":"Thermo Vanquish","COLUMN_NAME":"Phenomenex Kinetex C18 2.1 x 150 mm, 1.7 um","SOLVENT_A":"100% water; 0.1% formic acid","SOLVENT_B":"100% acetonitrile; 0.1% formic acid","FLOW_GRADIENT":"0-0.5 min 5% B, 0.5-1.1 min 5-95% B, 1.1-2.75 min hold at 95% B, 2.75-3 min 95-5% B, 3-5 min hold at 5% B.","FLOW_RATE":"450 uL/min","COLUMN_TEMPERATURE":"45","SAMPLE_INJECTION":"10 uL"},

"ANALYSIS":{"ANALYSIS_TYPE":"MS"},

"MS":{"INSTRUMENT_NAME":"Thermo Q Exactive Orbitrap","INSTRUMENT_TYPE":"Orbitrap","MS_TYPE":"ESI","ION_MODE":"POSITIVE","MS_COMMENTS":"Resolution 70,000, scan range 65-900 m/z, maximum injection time 200 ms, microscans 2, automatic gain control (AGC) 3 x 10^6 ions, source voltage 4.0 kV, capillary temperature 320 C, and sheath gas 45, auxiliary gas 15, and sweep gas 0 (all nitrogen). Data converted to mzXML using RawConverter. Metabolites were annotated and integrated using Maven in conjunction with the KEGG database."},

"MS_METABOLITE_DATA":{
"Units":"peak area",

"Data":[{"Metabolite":"L-leucine/isoleucine","1":"27215180","4":"28805310","7":"27636740","10":"34797230","13":"28219400","16":"43379600","19":"42851890","22":"36578500","25":"34454600","28":"19722300","31":"19903230","34":"21614110","2":"39810820","5":"34707530","8":"23546040","11":"33257360","14":"25885160","17":"21676690","20":"49355150","23":"53771920","26":"51423520","29":"34932040","32":"18866020","35":"34107500","3":"30690000","6":"46465420","9":"40980990","12":"35473880","15":"49648100","18":"42516120","21":"21356560","24":"57351630","27":"41288030","30":"25407170","33":"70818020"},{"Metabolite":"L-valine","1":"60218590","4":"51431160","7":"65579130","10":"57297020","13":"39206380","16":"68792990","19":"45958280","22":"71277590","25":"65534560","28":"35423210","31":"63660700","34":"41626540","2":"63525310","5":"72474180","8":"63148900","11":"59207280","14":"39259580","17":"58136700","20":"63855760","23":"83846550","26":"67293330","29":"55375090","32":"60518910","35":"54950160","3":"58044050","6":"74934730","9":"83326400","12":"77739060","15":"59787490","18":"86874860","21":"54153390","24":"90059580","27":"73366410","30":"53977200","33":"119463100"},{"Metabolite":"kynurenine","1":"72157.52","4":"142227.6","7":"189133.3","10":"186570.9","13":"130459.6","16":"198990.8","19":"128218.6","22":"160219.2","25":"102085.6","28":"122405.3","31":"108144.5","34":"313367.5","2":"82897.82","5":"190227.2","8":"125268.2","11":"194202.3","14":"110696.3","17":"113611","20":"258809.7","23":"130823.2","26":"105283.6","29":"170689.9","32":"76949.13","35":"249766.5","3":"116543.8","6":"149513.6","9":"154770.3","12":"237840.7","15":"213216.8","18":"186766.6","21":"177919.4","24":"169804.9","27":"204573.2","30":"128821.3","33":"216468.3"}],

"Metabolites":[{"Metabolite":"L-leucine/isoleucine","KEGG ID":"C00123","m/z":"132.102","medRt":"0.6674764"},{"Metabolite":"L-valine","KEGG ID":"C00183","m/z":"118.0865","medRt":"0.6581454"},{"Metabolite":"kynurenine","KEGG ID":"C00328","m/z":"209.0919","medRt":"0.6664889"}]
}

}